The following is offered as background information only and is not admitted to be prior art to the present invention.
Protein kinases (xe2x80x9cPKsxe2x80x9d) are enzymes that catalyze the phosphorylation of hydroxy groups on tyrosine, serine and threonine residues of proteins. The consequences of this seemingly simple activity are staggering; cell growth, differentiation and proliferation, i.e., virtually all aspects of cell life in one way or another depend on PK activity. Furthermore, abnormal PK activity has been related to a host of disorders, ranging from relatively non-life threatening diseases such as psoriasis to extremely virulent diseases such as glioblastoma (brain cancer).
The PKs can be conveniently broken down into two classes, the protein tyrosine kinases (PTKs) and the serine-threonine kinases (STKs).
One of the prime aspects of PTK activity is their involvement with growth factor receptors. Growth factor receptors are cell-surface proteins. When bound by a growth factor ligand, growth factor receptors are converted to an active form which interacts with proteins on the inner surface of a cell membrane. This leads to phosphorylation on tyrosine residues of the receptor and other proteins and to the formation inside the cell of complexes with a variety of cytoplasmic signaling molecules that, in turn, effect numerous cellular responses such as cell division (proliferation), cell differentiation, cell growth, expression of metabolic effects to the extracellular microenvironment, etc. For a more complete discussion, see Schlessinger and Ullrich, Neuron 9:303-391 (1992), which is incorporated by reference, including any drawings, as if fully set forth herein.
Growth factor receptors with PTK activity are known as receptor tyrosine kinases (xe2x80x9cRTKsxe2x80x9d). They comprise a large family of transmembrane receptors with diverse biological activity. At present, at least nineteen (19) distinct subfamilies of RTKs have been identified. An example of these is the subfamily designated the xe2x80x9cHERxe2x80x9d RTKs, which include EGFR (epithelial growth factor receptor), HER2, HER3 and HER4. These RTKs consist of an extracellular glycosylated ligand binding domain, a transmembrane domain and an intracellular cytoplasmic catalytic domain that can phosphorylate tyrosine residues on proteins.
Another RTK subfamily consists of insulin receptor (IR), insulin-like growth factor I receptor (IGF-1R) and insulin receptor related receptor (IRR). IR and IGF-1R interact with insulin, IGF-I and IGF-II to form a heterotetramer of two entirely extracellular glycosylated xcex1 subunits and two xcex2 subunits which cross the cell membrane and which contain the tyrosine kinase domain.
A third RTK subfamily is referred to as the platelet derived growth factor receptor (xe2x80x9cPDGFRxe2x80x9d) group, which includes PDGFRxcex1, PDGFRxcex2, CSFIR, c-kit and c-fms. These receptors consist of glycosylated extracellular domains composed of variable numbers of immunoglobin-like loops and an intracellular domain wherein the tyrosine kinase domain is interrupted by unrelated amino acid sequences.
Another group which, because of its similarity to the PDGFR subfamily, is sometimes subsumed into the later group is the fetus liver kinase (xe2x80x9cflkxe2x80x9d) receptor subfamily. This group is believed to be made of up of kinase insert domain-receptor fetal liver kinase-1 (KDR/FLK-1), flk-1R, flk-4 and fms-like tyrosine kinase 1 (flt-1).
A further member of the tyrosine kinase growth factor receptor family is the fibroblast growth factor (xe2x80x9cFGFxe2x80x9d) receptor subgroup. This group consists of four receptors, FGFR1-4, and seven ligands, FGF1-7. While not yet well defined, it appears that the receptors consist of a glycosylated extracellular domain containing a variable number of immunoglobin-like loops and an intracellular domain in which the tyrosine kinase sequence is interrupted by regions of unrelated amino acid sequences.
Still another member of the tyrosine kinase growth factor receptor family is the vascular endothelial growth factor (xe2x80x9cVEGFxe2x80x9d) receptor subgroup. VEGF is a dimeric glycoprotein similar to PDGF but has different biological functions and target cell specificity in vivo. In particular, VEGF is presently thought to play an essential role is vasculogenesis and angiogenesis.
Still another member of the tyrosine kinase growth factor receptor family is MET, often referred to as c-Met. c-Met is thought to play a role in priimary tumor growth and metastasis.
A more complete listing of the known RTK subfamilies is described in Plowman et al., DNandP, 7(6):334-339 (1994), which is incorporated by reference, including any drawings, as if fully set forth herein.
In addition to the RTKs, there also exists a family of entirely intracellular PTKs called xe2x80x9cnon-receptor tyrosine kinasesxe2x80x9d or xe2x80x9ccellular tyrosine kinases.xe2x80x9d This latter designation, abbreviated xe2x80x9cCTK,xe2x80x9d will be used herein. CTKs do not contain extracellular and transmembrane domains. At present, over 24 CTKs in 11 subfamilies (Src, Frk, Btk, Csk, Abl, Zap70, Fes, Fps, Fak, Jak and Ack) have been identified. The Src subfamily appear so far to be the largest group of CTKs and includes Src, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk. For a more detailed discussion of CTKs, see Bolen, Oncogene, 8:2025-2031 (1993), which is incorporated by reference, including any drawings, as if fully set forth herein.
The serine/threonine kinases, STKs, like the CTKs, are predominantly intracellular although there are a few receptor kinases of the STK type. STKs are the most common of the cytosolic kinases; i.e., kinases that perform their function in that part of the cytoplasm other than the cytoplasmic organelles and cytoskelton. The cytosol is the region within the cell where much of the cell""s intermediary metabolic and biosynthetic activity occurs; e.g., it is in the cytosol that proteins are synthesized on ribosomes.
RTKs, CTKs and STKs have all been implicated in a host of pathogenic conditions including, significantly, cancer. Other pathogenic conditions which have been associated with PTKs include, without limitation, psoriasis, hepatic cirrhosis, diabetes, angiogenesis, restenosis, ocular diseases, rheumatoid arthritis and other inflammatory disorders, immunological disorders such as autoimmune disease, cardiovascular disease such as atherosclerosis and a variety of renal disorders.
With regard to cancer, two of the major hypotheses advanced to explain the excessive cellular proliferation that drives tumor development relate to functions known to be PK regulated. That is, it has been suggested that malignant cell growth results from a breakdown in the mechanisms that control cell division and/or differentiation. It has been shown that the protein products of a number of proto-oncogenes are involved in the signal transduction pathways that regulate cell growth and differentiation. These protein products of proto-oncogenes include the extracellular growth factors, transmembrane growth factor PTK receptors (RTKs), cytoplasmic PTKs (CTKs) and cytosolic STKs, discussed above.
In view of the apparent link between PK-related cellular activities and wide variety of human disorders, it is no surprise that a great deal of effort is being expended in an attempt to identify ways to modulate PK activity. Some of these have involved biomimetic approaches using large molecules patterned on those involved in the actual cellular processes (e.g., mutant ligands (U.S. application Ser. No. 4,966,849); soluble receptors and antibodies (Application No. WO 94/10202, Kendall and Thomas, Proc. Nat""l Acad. Sci., 90:10705-10709 (1994), Kim, et al., Nature, 362:841-844 (1993)); RNA ligands (Jelinek, et al., Biochemistry, 33:10450-56); Takano, et al., Mol. Bio. Cell, 4:358A (1993); Kinsella, et al., Exp. Cell Res., 199:56-62 (1992); Wright, et al., J. Cellular Phys., 152:448-57) and tyrosine kinase inhibitors (WO 94/03427; WO 92/21660; WO 91/15495; WO 94/14808; U.S. Pat. No. 5,330,992; Mariani, et al., Proc. Am. Assoc. Cancer Res., 35:2268 (1994)).
In addition to the above, attempts have been made to identify small molecules which act as PK inhibitors. For example, bis-monocylic, bicyclic and heterocyclic aryl compounds (PCT WO 92/20642), vinylene-azaindole derivatives (PCT WO 94/14808) and 1-cyclopropyl-4-pyridylquinolones (U.S. Pat. No. 5,330,992) have been described as tyrosine kinase inhibitors. Styryl compounds (U.S. Pat. No. 5,217,999), styryl-substituted pyridyl compounds (U.S. Pat. No. 5,302,606), quinazoline derivatives (EP Application No. 0 566 266 A1), selenaindoles and selenides (PCT WO 94/03427), tricyclic polyhydroxylic compounds (PCT WO 92/21660) and benzylphosphonic acid compounds (PCT WO 91/15495) have all been described as PTK inhibitors useful in the treatment of cancer.
A family of novel pyrrole-substituted 2-indolinone compounds have been discovered which exhibit PK modulating ability and have a salutary effect against disorders related to abnormal PK activity (U.S. Pat. No. 5,792,783; U.S. application Ser. No. 09/322,297). It has been demonstrated that this family of compounds modulates the catalytic activity of receptor tyrosine kinases (RTKs), non-receptor protein tyrosine kinases (CTKs) and serine/threonine protein kinases (STKs).
For example, the catalytic activity of RTKs such as, without limitation, EGF, MET, HER2, HER3, HER4, IR, IGF-1R, IRR, PDGFRxcex1, PDGFRxcex2, CSFIR, C-Kit, C-fms, Flk-1R, Flk4, KDR/Flk-1, Flt-1, FGFR-1R, FGFR-2R, FGFR-3R and FGFR-4R may be modulated with pyrrole-substituted 2-indolinone compounds, in particular Met may be modulated. By affecting the catalytic activity of RTKs, CTKs and/or STKs, such compounds can interfere with the signals transduced by such proteins. This application also encompasses affecting the catalytic activity of protein kinases disclosed in U.S. application Ser. Nos. 09/233,857; 09/291,417; 60/149,005 and 60/136,503, and PCT Application No. PCT/US99/13533, the entire disclosures of which are hereby incorporated by reference.
In one aspect, the present invention relates to compounds having the following chemical structure (Formula I). 
wherein:
R1 is an aryl or heteroaryl substituent, optionally substituted by one or more substituent selected from the group consisting of halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, xe2x80x94OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkyl further substituted by one or more of R2, lower alkenyl, lower alkenyl further substituted by one or more of R2, lower alkenyl, lower alkenyl, lower alkenyl further substituted by one or more of R2, cycloalkyl, cycloalkyl further substituted by one or more of R2, a heterocyclic ring, a heterocyclic ring further substituted by one or more of R2, aryl and aryl further substituted by one or more of R2;
R2 is selected from the group consisting of hydrogen, halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, xe2x80x94OCOR6, CONR6R7, xe2x80x94RNCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, a heterocyclic ring and aryl;
R3 is selected from the group consisting of hydrogen, halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, xe2x80x94OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, a heterocyclic ring, a heterocyclic ring further substituted by one or more of R2, and aryl, wherein said lower alkyl is further substituted by xe2x80x94CONR6R7, NR6R7, xe2x80x94SO2R6, xe2x80x94R6NSO2R, or xe2x80x94SO2NR6R7;
R4 is selected from the group consisting of hydrogen, halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, xe2x80x94OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, a heterocyclic ring and aryl, wherein said lower alkyl is further substituted by xe2x80x94CONR6R7, NR6R7, xe2x80x94SO2R6, xe2x80x94R6NSO2R, or SO2NR6R7;
R5 is selected from the group consisting of hydrogen, halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, xe2x80x94OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, a heterocyclic ring and aryl;
provided that no more than one of R3, R4, or R5 is hydrogen;
R3 and R4 or R4 and R5 may be linked together to form a 4-, 5-, 6- or 7-membered ring optionally containing one or more hetero atoms selected from the group consisting of O, N, S, SO and SO2, which may contain 1 or 2 double bonds and may be further substituted by one or more of xe2x80x94(CH2)nxe2x80x94NR6R7, xe2x80x94(CH2)nxe2x80x94CR6R7, xe2x80x94(CH2)nxe2x80x94C(O)xe2x80x94(CH2)nxe2x80x94NR6R7, xe2x80x94(CH2)nSO2R6R7, xe2x80x94(CH2)nNSO2R6R7 or xe2x80x94(CH2)nxe2x80x94C(O)xe2x80x94R6, wherein n is 0-4;
R6 and R7 are independently selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, a heterocyclic ring and aryl;
wherein lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, the heteocyclic ring or aryl may be further substituted by one or more of (i) xe2x80x94NR12R13, (ii) hydroxy, (iii) halo, (iv) a heterocyclic ring, (v) lower alkyl, (vi) xe2x80x94C(O)xe2x80x94NR12R13, (vii) xe2x80x94OR12, (viii) xe2x80x94SO2R12R13, or (ix) xe2x80x94COR6;
wherein said heterocyclic ring (iv) may be further substituted by one or more of lower alkyl, xe2x80x94COR12, xe2x80x94NR12COR13, halogen, xe2x80x94OR12, CX3, xe2x80x94C(O)NR12R13, xe2x80x94SO2R12R13, or xe2x80x94SO2NR12R13,
or R6 and R7 may be linked together to form a 4, 5- or 6-membered ring, optionally containing a hetero atom selected from the group consisting of N, O, S and SO2, which may be further substituted by xe2x80x94CONR12R13, lower alkyl, hydroxy, xe2x80x94(CH2)nxe2x80x94NR12R13, xe2x80x94(CH2)n-heterocycle, xe2x80x94(CH2)nxe2x80x94C(O)xe2x80x94N12R13, xe2x80x94(CH2)nSO2R12R13, or xe2x80x94(CH2)nNSO2R12R13, wherein said heterocycle may be further substituted by halo, lower alkyl, xe2x80x94COR12, hydroxy, xe2x80x94C(O)xe2x80x94NR12R13, xe2x80x94OR12, xe2x80x94SO2R12R13, or xe2x80x94SO2NR12R13;
X is fluorine, chlorine, bromine or iodine;
R12 is selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, xe2x80x94(CH2)n-cycloalkyl, xe2x80x94(CH2)n-heterocycle, and aryl;
R13 is selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, xe2x80x94(CH2)n-cycloalkyl, xe2x80x94(CH2)n-heterocycle, and aryl;
or R12 and R13 may be linked together to form a 4-, 5- or 6-membered ring optionally containing one or more hetero atoms selected from the group consisting of O, N, S, SO and SO2, which may contain 1 or 2 double bonds;
or a pharmaceutically acceptable salt thereof.
In a preferred aspect, R3 and R4 or R4 and R5 may be linked together to form a ring.
In a particularly preferred aspect, R3 and R4 or R4 and R5 are linked together to form a ring, the ring together with pyrrole is selected from the group consisting of: 
In another preferred aspect, R3 may be excluded when both R4 and R5 are hydrogen. In still a further preferred aspect, R1 is aryl.
A preferred aspect of the invention relates to compounds of Formula II 
wherein each R8 is independently halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94RNCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkyl further substituted by one or more of R2, lower alkenyl, lower alkenyl further substituted by one or more of R2, lower alkenyl, lower alkenyl further substituted by one or more of R2, cycloalkyl, cycloalkyl further substituted by one or more of R2, a heterocyclic ring, a heterocylic ring further substituted by one or more of R2, aryl and aryl further substituted by one or more of R2;
R2 is selected from the group consisting of hydrogen, halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94R12NCOR13, xe2x80x94NR6R7, xe2x80x94RNC(O)R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, a heterocyclic ring and aryl;
R6 and R7 are independently selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, a heterocyclic ring and aryl;
wherein lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, the heteocyclic ring or aryl may be further substituted by one or more of (i) xe2x80x94NR12R13, (ii) hydroxy, (iii) halo, (iv) a heterocyclic ring, (v) lower alkyl, (vi) xe2x80x94C(O)xe2x80x94NR12R13, (vii) xe2x80x94OR12, (viii) xe2x80x94SO2R12R13, or (ix) xe2x80x94COR6;
wherein said heterocyclic ring (iv) may be further substituted by one or more of lower alkyl, xe2x80x94COR12, xe2x80x94NR12COR13, halogen, xe2x80x94OR12, CX3, xe2x80x94C(O)NR12R13, xe2x80x94SO2R12R13, or xe2x80x94SO2NR12R13,
or R6 and R7 may be linked together to form a 4, 5- or 6-membered ring, optionally containing a hetero atom selected from the group consisting of N, O, S and SO2, which may be further substituted by xe2x80x94CONR12R13, lower alkyl, hydroxy, xe2x80x94(CH2)nxe2x80x94NR12R13, xe2x80x94(CH2)n-heterocycle, xe2x80x94(CH2)nxe2x80x94C(O)xe2x80x94NR12R13, xe2x80x94(CH2)nSO2R12R13, or xe2x80x94(CH2)nNSO2R12R13, wherein said heterocycle may be further substituted by halo, lower alkyl, xe2x80x94COR12, hydroxy, xe2x80x94C(O)xe2x80x94NR12R13, xe2x80x94OR12, xe2x80x94SO2R12R13, or xe2x80x94SO2NR12R13;
X is fluorine, chlorine, bromine or iodine;
R12 is selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, xe2x80x94(CH2)n-cycloalkyl, xe2x80x94(CH2)n-heterocycle, and aryl;
R13 is selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, xe2x80x94(CH2)n-cycloalkyl, xe2x80x94(CH2)n-heterocycle, and aryl;
or R12 and R13 may be linked together to form a 4-, 5- or 6-membered ring optionally containing one or more hetero atoms selected from the group consisting of O, N, S, SO and SO2, which may contain 1 or 2 double bonds and may be further substituted by halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94R12NCOR13, xe2x80x94NR6R7, xe2x80x94R6NC(O)R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94RNSO2R7, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkenyl, cycloalkyl, a heterocyclic ring and aryl;
R9 is selected from the group consisting of halogen, xe2x80x94CX3, lower alkyl, cycloalkyl, a heterocyclic ring and aryl, each of which may be further substituted by halogen, xe2x80x94OR6, xe2x80x94COOR6, xe2x80x94OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CX3, xe2x80x94SR6, SOR6, SO2R6, xe2x80x94SO2OR6, SO2NR6R7, or xe2x80x94R6NSO2R7; and
m is 1 or 2,
or a pharmaceutically acceptable salt thereof.
Another preferred aspect of the invention relates to compounds of Formula III, 
wherein each R8 is independently halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94RNCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkyl further substituted by one or more of R2, lower alkenyl, lower alkenyl further substituted by one or more of R2, lower alkynyl, lower alkynyl further substituted by one or more of R2, cycloalkyl, cycloalkyl further substituted by one or more of R2, a heterocyclic ring, a heterocyclic ring further substituted by one or more of R2, aryl and aryl further substituted by one or more of R2;
R2 is selected from the group consisting of hydrogen, halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR R7, xe2x80x94R6COR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, xe2x80x94R6NSO2R7, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, a heterocyclic ring and aryl;
R6 and R7 are independently selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, a heterocyclic ring and aryl;
wherein lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, the heteocyclic ring or aryl may be further substituted by one or more of (i) xe2x80x94NR12R13, (ii) hydroxy, (iii) halo, (iv) a heterocyclic ring, (v) lower alkyl, (vi) xe2x80x94C(O)xe2x80x94NR12R13, (vii) xe2x80x94OR12, (viii) xe2x80x94SO2R12R13, or (ix) xe2x80x94COR6;
wherein said heterocyclic ring (iv) may be further substituted by one or more of lower alkyl, xe2x80x94COR12, xe2x80x94NR12COR13, halogen, OR12, CX3, xe2x80x94C(O)NR12R13, xe2x80x94SO2R12R13, or xe2x80x94SO2NR12R13,
or R6 and R7 may be linked together to form a 4, 5- or 6-membered ring, optionally containing a hetero atom selected from the group consisting of N, O, S and SO2, which may be further substituted by xe2x80x94CONR12R13, lower alkyl, hydroxy, xe2x80x94(CH2)nxe2x80x94NR12R13, xe2x80x94(CH2)n-heterocycle, xe2x80x94(CH2)nxe2x80x94C(O)xe2x80x94NR12R13, xe2x80x94(CH2)nSO2R12R13, or xe2x80x94(CH2)nNSO2R12R13, wherein said heterocycle may be further substituted by halo, lower alkyl, xe2x80x94COR12, hydroxy, xe2x80x94C(O)xe2x80x94NR12R13, xe2x80x94OR12, xe2x80x94SO2R12R13, or xe2x80x94SO2NR12R13;
X is fluorine, chlorine, bromine or iodine;
R10 is H, lower alkyl, lower alkyl substituted with one or more of R2, xe2x80x94(CH2)nNR6R7, xe2x80x94CONR6R7, xe2x80x94SO2NR6R7, xe2x80x94(CH2)nxe2x80x94SR6, xe2x80x94(CH2)nxe2x80x94SOR6, xe2x80x94(CH2)nxe2x80x94SO2NR6R7, or xe2x80x94(CH2)nxe2x80x94OR6;
R11 is H, lower alkyl, lower alkyl substituted with one or more of R2, xe2x80x94(CH2)nNR6R7, xe2x80x94CONR6R7, xe2x80x94SO2NR6R7, xe2x80x94(CH2)nxe2x80x94SR6, xe2x80x94(CH2)nxe2x80x94SOR6, xe2x80x94(CH2)nxe2x80x94SO2NR6R7, or xe2x80x94(CH2)nxe2x80x94OR6;
R12 is selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkynyl, xe2x80x94(CH2)n-cycloalkyl, xe2x80x94(CH2)n-heterocycle, and aryl;
R13 is selected from the group consisting of hydrogen, xe2x80x94CX3, perfluoroalkyl, lower alkyl, lower alkenyl, lower alkynyl, xe2x80x94(CH2)n-cycloalkyl, xe2x80x94(CH2)n-heterocycle, and aryl;
or R12 and R13 may be linked together to form a 4-, 5- or 6-membered ring optionally containing one or more hetero atoms selected from the group consisting of O, N, S, SO and SO2, which may contain 1 or 2 double bonds; and
wherein  is a single or double bond; and
n is 0-4,
or a pharmaceutically acceptable salt thereof.
The compounds presented herein are exemplary only and are not to be construed as limiting the scope of this invention in any manner.
The chemical formulae referred to herein may exhibit the phenomena of tautomerism and structural isomerism. For example, the compounds described herein may adopt an E or a Z configuration about the double bond connecting the 2-indolinone moiety to the pyrrole moiety or they may be a mixture of E and Z. This invention encompasses any tautomeric or structural isomeric form and mixtures thereof which possess the ability to modulate RTK, CTK and/or STK activity and is not limited to any one tautomeric or structural isomeric form.
In addition, the formulae referred to herein may also exhibit stereoisomerism, in which such compounds may adopt an R or S configuration at chiral centers. Thus, this invention also encompasses any stereoisomeric form, their corresponding enantiomers (d- and 1- or (+) and (xe2x88x92) isomers) and diastereomers thereof, and mixtures thereof, which possess the ability to modulate RTK, CTK and/or STK activity and is not limited to any one stereoisomeric form.
Table 1 shows the chemical structures of the preferred compounds of the invention.
In a second aspect, this inventions is directed to a pharmaceutical composition comprising one or more compounds of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
Another aspect of this invention relates to a method for the modulation of the catalytic activity of a PK by contacting a PK with a compound of this invention or a physiologically acceptable salt thereof.
A further aspect of this invention is that the modulation of the catalytic activity of PKs using a compound of this invention may be carried out in vitro or in vivo.
A still further aspect of this invention is that the protein kinase whose catalytic activity is being modulated by a compound of this invention is selected from the group consisting of receptor protein tyrosine kinases, cellular tyrosine kinases and serine-threonine kinases.
It is an aspect of this invention that the receptor tyrosine protein kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of EGF, HER2, HER3, HER4, IR, IGF-1R, IRR, PDGFRxcex1, PDGFRxcex2, CSFIR, C-Kit, C-fms, Flk-1R, Flk4, KDR/Flk-1, Flt-1, FGFR-1R, FGFR-2R, FGFR-3R, FGFR-4R, MET, DDR-1 and DDR-2.
In addition, it is an aspect of this invention that the cellular tyrosine kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of Src, Frk, Btk, Csk, Abl, ZAP70, Fes/Fps, Fak, Jak, Ack, Yes, Lyn, Lck, Blk, Hck, Fgr and Yrk.
Another aspect of this invention is that the serine-threonine protein kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of CDK2, Raf, NEK and BUB1.
Another aspect of this inventions relates to a method for treating or preventing a protein kinase related disorder in an organism comprising administering a therapeutically effective amount of a compound or a salt thereof Formula I to an organism.
It is an aspect of this invention that the above-referenced protein kinase related disorder is selected from the group consisting of a receptor protein tyrosine kinase related disorder, a cellular tyrosine kinase disorder and a serine-threonine kinase related disorder.
In yet another aspect of this invention, the above referenced protein kinase related disorder is selected from the group consisting of a Met related disorder, a PDGFR related disorder, an IGFR related disorder and a flk related disorder.
The above referenced protein kinase related disorders include by way of example and not limitation, cancers such as lung cancer, NSCLC (non small cell ling cancer), bone cancer, pancreatic cancer, skin cancer, cancer of the head and neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, gynecologic tumors (e.g., uterine sarcomas, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina or carcinoma of the vulva), Hodgkin""s Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system (e.g., cancer of the thyroid, parathyroid or adrenal glands), sarcomas of soft tissues, cancer of the urethra, cancer of the penis, prostate cancer, chronic or acute leukemia, solid tumors of childhood, lynphocytic lymphomas, cancer of the bladder, cancer of the kidney or uerter (e.g., renal cell carcinoma, carcinoma of the renal pelvis), pediatric malignancy, neoplasms of the central nervous system (e.g., primary CNS lymphoma, spinal axis tumors, brain stem glima or pituitary adenomas), Barrett""s esophagus (pre-malignant syndrome), neoplastic cutaneous disease, psoriasis, mycoses fungoides and benign prostatic hypertrophy, diabetes related diseases such as diabetic retinopathy, retinal ischemia and retinal neovascularization, hepatic cirrhosis, cardiovascular disease such as atherosclerosis, immunological disease such as autoimmune disease and renal disease. Preferably, the disease is cancer such as acute myeloid leukemia and colorectal cancer.
The above referenced protein kinase related disorder also includes disorders selected from the group consisting of diabetes, a hyper-proliferation disorder, hyperproliferative disorders of the kidney, von Hippel-Lindau disease, restenosis, fibrosis, psoriasis, osteoarthritis, rheumatoid arthritis, an inflammatory disorder and angiogenesis in yet another aspect of this invention.
Additional disorders which may be treated or prevented using the compounds of this invention are immunological disorders such as autoimmune diseases (e.g., AIDS, lupus, etc.) and cardiovascular disorders such as atherosclerosis.
It is an aspect of this invention that the protein kinase related disorder being treated or prevented by administration of a compound of this invention is a CDK2 related disorder.
The organism in which the protein kinase related disorder is being treated or prevented is a human being in yet another aspect of this invention.
It is as aspect of this invention that a chemical compound that modulates the catalytic activity of a protein kinase may be identified by contacting cells expressing said protein kinase with a compound or a salt thereof of Formula I and then monitoring said cells for an effect.
The above-referenced effect is selected from a change or an absence of change in a cell phenotype, a change or absence of change in the catalytic activity of said protein kinase or a change or absence of change in the interaction of said protein kinase with a natural binding partner in a final aspect of this invention.
It is also an aspect of this invention that a compound described herein, or its salt, might be combined with other chemotherapeutic agents for the treatment of the diseases and disorders discussed above. For instance, a compound or salt of this invention might be combined with alkylating agents such as fluorouracil (5-FU) alone or in further combination with leukovorin; or other alkylating agents such as, without limitation, other pyrimidine analogs such as UFT, capecitabine, gemcitabine and cytarabine, the alkyl sulfonates, e.g., busulfan (used in the treatment of chronic granulocytic leukemia), improsulfan and piposulfan; aziridines, e.g., benzodepa, carboquone, meturedepa and uredepa; ethyleneimines and methylmelamines, e.g., altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolmelamine; and the nitrogen mustards, e.g., chlorambucil (used in the treatment of chronic lymphocytic leukemia, primary macroglobulinemia and non-Hodgkin""s lymphoma), cyclophosphamide (used in the treatment of Hodgkin""s disease, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, Wilm""s tumor and rhabdomyosarcoma), estramustine, ifosfamide, novembrichin, prednimustine and uracil mustard (used in the treatment of primary thrombocytosis, non-Hodgkin""s lymphoma, Hodgkin""s disease and ovarian cancer); and triazines, e.g., dacarbazine (used in the treatment of soft tissue sarcoma).
Likewise a compound or salt of this invention might be expected to have a beneficial effect in combination with other antimetabolite chemotherapeutic agents such as, without limitation, folic acid analogs, e.g. methotrexate (used in the treatment of acute lymphocytic leukemia, choriocarcinoma, mycosis fungiodes breast cancer, head and neck cancer and osteogenic sarcoma) and pteropterin; and the purine analogs such as mercaptopurine and thioguanine which find use in the treatment of acute granulocytic, acute lymphocytic and chronic granulocytic leukemias.
A compound or salt of this invention might also be expected to prove efficacious in combination with natural product based chemotherapeutic agents such as, without limitation, the vinca alkaloids, e.g., vinblastin (used in the treatment of breast and testicular cancer), vincristine and vindesine; the epipodophylotoxins, e.g., etoposide and teniposide, both of which are useful in the treatment of testicular cancer and Kaposi""s sarcoma; the antibiotic chemotherapeutic agents, e.g., daunorubicin, doxorubicin, epirubicin, mitomycin (used to treat stomach, cervix, colon, breast, bladder and pancreatic cancer), dactinomycin, temozolomide, plicamycin, bleomycin (used in the treatment of skin, esophagus and genitourinary tract cancer); and the enzymatic chemotherapeutic agents such as L-asparaginase.
In addition to the above, a compound or salt of this invention might be expected to have a beneficial effect used in combination with the platinum coordination complexes (cisplatin, etc.); substituted ureas such as hydroxyurea; methylhydrazine derivatives, e.g., procarbazine; adrenocortical suppressants, e.g., mitotane, aminoglutethimide; and hormone and hormone antagonists such as the adrenocorticosteriods (e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate); estrogens (e.g., diethylstilbesterol); antiestrogens such as tamoxifen; androgens, e.g., testosterone propionate; and aromatase inhibitors (such as anastrozole.
Finally, the combination of a compound of this invention might be expected to be particularly effective in combination with mitoxantrone or paclitaxel for the treatment of solid tumor cancers or leukemias such as, without limitation, acute myelogenous (non-lymphocytic) leukemia.
The above method can be carried out in combination with a chemotherapeutic agent selected from the group consisting of mitotic inhibitors, alkylating agetns, antimetabolites, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, antiangiogenic agents such as MMP-2, MMP-9 and COX-2 inhibitors, and anti-androgens.
Examples of useful COX-II inhibitors include Vioxx(trademark), CELEBREX(trademark) (alecoxib), valdecoxib, paracoxib, rofecoxib, and Cox 189. Examples of useful matrix metalloproteinase inhibitors are described in WO 96/33172 (published Oct. 24, 1996), WO 96/27583 (published Mar. 7, 1996), European Patent Application No. 97304971.1 (filed Jul. 8, 1997), European Patent Application No. 99308617.2 (filed Oct. 29, 1999), WO 98/07697 (published Feb. 26, 1998), WO 98/03516 (published Jan. 29, 1998), WO 98/34918 (published Aug. 13, 1998), WO 98/34915 (published Aug. 13, 1998), WO 98/33768 (published Aug. 6, 1998), WO 98/30566 (published Jul. 16, 1998), European Patent Publication 606,046 (published Jul. 13, 1994), European Patent Publication 931,788 (published Jul. 28, 1999), WO 90/05719 (published May 31, 1990), WO 99/52910 (published Oct. 21, 1999), WO 99/52889 (published Oct. 21, 1999), WO 99/29667 (published Jun. 17, 1999), PCT International Application No. PCT/IB98/01113 (filed Jul. 21, 1998), European Patent Application No. 99302232.1 (filed Mar. 25, 1999), Great Britain patent application number 9912961.1 (filed Jun. 3, 1999), U.S. Provisional Application No. 60/148,464 (filed Aug. 12, 1999), U.S. Pat. No. 5,863,949 (issued Jan. 26, 1999), U.S. Pat. No. 5,861,510 (issued Jan. 19, 1999), and European Patent Publication 780,386 (published Jun. 25, 1997), all of which are incorporated herein in their entireties by reference. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity inhibiting MMP-1. More preferred, are those that selectively inhibit MMP-2 and/or MMP-9 relative to the other matrix-metalloproteinases (i.e. MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).
Some specific examples of MMP inhibitors useful in the present invention are AG-3340, RO 32-3555, RS 13-0830, and the compounds recited in the following list:
3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl)-amino]-propionic acid; 3-exo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic acid hydroxyamide; (2R,3R) 1-[4-(2-chloro-4-fluoro-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide; 4-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylic acid hydroxyamide; 3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl)-amino]-propionic acid; 4-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylic acid hydroxyamide; (R) 3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-3-carboxylic acid hydroxyamide; (2R,3R) 1-[4-(4-fluoro-2-methyl-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide; 3-[[(4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-ethyl)-amino]-propionic acid; 3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(4-hydroxycarbamoyl-tetrahydro-pyran-4-yl)-amino]-propionic acid; 3-exo-3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic acid hydroxyamide; 3-endo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic acid hydroxyamide; and (R) 3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-furan-3-carboxylic acid hydroxyamide; and pharmaceutically acceptable salts and solvates of said compounds.
Other anti-angiogenesis agents, including other COX-II inhibitors and other MMP inhibitors, can also be used in the present invention.
A compound of Formula (I) can also be used with signal transduction inhibitors, such as agents that can inhibit EGFR (epidermal growth factor receptor) responses, such as EGFR antibodies, EGF antibodies, and molecules that are EGFR inhibitors; VEGF (vascular endothelial growth factor) inhibitors; and erbB2 receptor inhibitors, such as organic molecules or antibodies that bind to the erbB2 receptor, for example, HERCEPTIN.(trademark). (Genentech, Inc. of South San Francisco, Calif., USA). EGFR inhibitors are described in, for example in WO 95/19970 (published Jul. 27, 1995), WO 98/14451 (published Apr. 9, 1998), WO 98/02434 (published Jan. 22, 1998), and U.S. Pat. No. 5,747,498 (issued May 5, 1998), and such substances can be used in the present invention as described herein.
EGFR-inhibiting agents include, but are not limited to, the monoclonal antibodies C225 and anti-EGFR 22Mab (ImClone Systems Incorporated of New York, N.Y., USA), the compounds ZD-1839 (AstraZeneca), BIBX-1382 (Boehringer Ingelheim), MDX-447 (Medarex Inc. of Annandale, N.J., USA), and OLX-103 (Merck and Co. of Whitehouse Station, N.J., USA), VRCTC-310 (Ventech Research) and EGF fusion toxin (Seragen Inc. of Hopkinton, Mass.).
These and other EGFR-inhibiting agents can be used in the present invention.
VEGF inhibitors, for example SU-5416, SU 11248, SU-6668 (Sugen Inc. of South San Francisco, Calif., USA), can also be combined with a compound of Formula (I). VEGF inhibitors are described in, for example in WO 99/24440 (published May 20, 1999), PCT International Application PCT/IB99/00797 (filed May 3, 1999), in WO 95/21613 (published Aug. 17, 1995), WO 99/61422 (published Dec. 2, 1999), U.S. Pat. No. 5,834,504 (issued Nov. 10, 1998), WO 01/60814, WO 98/50356 (published Nov. 12, 1998), U.S. Pat. No. 5,883,113 (issued Mar. 16, 1999), U.S. Pat. No. 5,886,020 (issued Mar. 23, 1999), U.S. Pat. No. 5,792,783 (issued Aug. 11, 1998), WO 99/10349 (published Mar. 4, 1999), WO 97/32856 (published Sep. 12, 1997), WO 97/22596 (published Jun. 26, 1997), WO 98/54093 (published Dec. 3, 1998), WO 98/02438 (published Jan. 22, 1998), WO 99/16755 (published Apr. 8, 1999), and WO 98/02437 (published Jan. 22, 1998), all of which are incorporated herein in their entireties by reference. Other examples of some specific VEGF inhibitors useful in the present invention are IM862 (Cytran Inc. of Kirkland, Wash., USA); anti-VEGF monoclonal antibody of Genentech, Inc. of South San Francisco, Calif.; and angiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colo.) and Chiron (Emeryville, Calif.). These and other VEGF inhibitors can be used in the present invention as described herein.
ErbB2 receptor inhibitors, such as GW-282974 (Glaxo Wellcome plc), and the monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc. of The Woodlands, Tex., USA) and 2B-1 (Chiron), can furthermore be combined with a compound of Formula (I) for example those indicated in WO 98/02434 (published Jan. 22, 1998), WO 99/35146 (published Jul. 15, 1999), WO 99/35132 (published Jul. 15, 1999), WO 98/02437 (published Jan. 22, 1998), WO 97/13760 (published Apr. 17, 1997), WO 95/19970 (published Jul. 27, 1995), U.S. Pat. No. 5,587,458 (issued Dec. 24, 1996), and U.S. Pat. No. 5,877,305 (issued Mar. 2, 1999), which are all hereby incorporated herein in their entireties by reference. ErbB2 receptor inhibitors useful in the present invention are also described in U.S. Provisional Application No. 60/117,341, filed Jan. 27, 1999, and in U.S. Provisional Application No. 60/117,346, filed Jan. 27, 1999, both of which are incorporated in their entireties herein by reference. The erbB2 receptor inhibitor compounds and substance described in the aforementioned PCT applications, U.S. patents, and U.S. provisional applications, as well as other compounds and substances that inhibit the erbB2 receptor, can be used with a compound of Formula (I), in accordance with the present invention.
A compound of Formula (I) can also be used with other agents useful in treating cancer, including, but not limited to, agents capable of enhancing antitumor immune responses, such as CTLA4 (cytotoxic lymphocite antigen 4) antibodies, and other agents capable of blocking CTLA4; and anti-proliferative agents such as other farnesyl protein transferase inhibitors, for example the farnesyl protein transferase inhibitors described in the references cited in the xe2x80x9cBackgroundxe2x80x9d section, of U.S. Pat. No. 6,258,824 B1. Specific CTLA4 antibodies that can be used in the present invention include those described in U.S. Provisional Application 60/113,647 (filed Dec. 23, 1998) which is incorporated by reference in its entirety, however other CTLA4 antibodies can be used in the present invention.
The above method can be also be carried out in combination with radiation therapy, wherein the amount of a compound of Formula (I) in combination with the radiation therapy effective in treating the above diseases.
Techniques for administering radiation therapy are known in the art, and these techniques can be used in the combination therapy described herein. The administration of the compound of the invention in this combination therapy can be determined as described herein.
Another aspect of the invention is directed to the use of a compound of Formula (I) in the preparation of a medicament, which is useful in the treatment of a disease mediated by abnormal Met kinase activity.
The terms xe2x80x9cindolinonexe2x80x9d, xe2x80x9c2-indolinonexe2x80x9d and xe2x80x9cindolin-2-onexe2x80x9d are used interchangeably herein to refer to a molecule having the chemical structure: 
xe2x80x9cPyrrolexe2x80x9d refers to a molecule having the chemical structure: 
xe2x80x9cPyrrole-substituted 2-indolinonexe2x80x9d and xe2x80x9c3-pyrrolidinyl-2-indolinonexe2x80x9d are used interchangeably herein to refer to a chemical compound having the general structure shown in Formula I. 
xe2x80x9cPharmaceutically acceptable saltxe2x80x9d or xe2x80x9cpharmaceutically acceptable salt thereofxe2x80x9d refer to those salts which retain the biological effectiveness and properties of the free bases and which are obtained by reaction with inorganic or organic acids, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, acetic acid, benzenesulfonic acid (besylate), benzoic acid, camphorsulfonic acid, citric acid, fumaric acid, gluconic acid, glutamic acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, mucic acid, pamoic acid, pantothenic acid, succinic acid, tartaric acid, and the like.
A xe2x80x9cpharmaceutical compositionxe2x80x9d refers to a mixture of one or more of the compounds described herein, or physiologically acceptable salts thereof, with other chemical components, such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.
As used herein, a xe2x80x9cphysiologically acceptable carrierxe2x80x9d refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.
An xe2x80x9cexcipientxe2x80x9d refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like.
xe2x80x9cAlkylxe2x80x9d refers to a saturated aliphatic hydrocarbon including straight chain, branched chain or cyclic groups. Preferably, the alkyl group has 1 to 20 carbon atoms (whenever a numerical range; e.g., xe2x80x9c1-20xe2x80x9d, is stated herein, it means that the group, in this case the alkyl group, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc. up to and including 20 carbon atoms). More preferably, it is a medium size alkyl having 1 to 10 carbon atoms. Most preferably, it is a lower alkyl having 1 to 4 carbon atoms. The alkyl group may be substituted or unsubstituted. When substituted, the substituent group(s) is preferably one or more individually selected from halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, thiocarbonyl, xe2x80x94R(NSO2R7, perfluoroalkyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, silyl, ammonium, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycle, heteroaryl and aryl.
xe2x80x9cAlkenylxe2x80x9d refers to an aliphatic hydrocarbon having at least one carbon-carbon double bond, including straight chain, branched chain or cyclic groups having at least one carbon-carbon double bond. Preferably, the alkenyl group has 2 to 20 carbon atoms (whenever a numerical range; e.g., xe2x80x9c2-20xe2x80x9d, is stated herein, it means that the group, in this case the alkenyl group, may contain 2 carbon atoms, 3 carbon atoms, etc. up to and including 20 carbon atoms). More preferably, it is a medium size alkenyl having 2 to 10 carbon atoms. Most preferably, it is a lower alkenyl having 2 to 6 carbon atoms. The alkenyl group may be substituted or unsubstituted.
When substituted, the substituent group(s) is preferably one or more individually selected from halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, thiocarbonyl, xe2x80x94R6NSO2R7, perfluoroalkyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, silyl, ammonium, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycle, heteroaryl and aryl.
xe2x80x9cAlkynylxe2x80x9d refers to an aliphatic hydrocarbon having at least one carbon-carbon triple bond, including straight chain, branched chain or cyclic groups having at least one carbon-carbon triple bond. Preferably, the alkenyl group has 2 to 20 carbon atoms (whenever a numerical range; e.g., xe2x80x9c2-20xe2x80x9d, is stated herein, it means that the group, in this case the alkynyl group, may contain 2 carbon atoms, 3 carbon atoms, etc. up to and including 20 carbon atoms). More preferably, it is a medium size alkynyl having 2 to 10 carbon atoms. Most preferably, it is a lower alkynyl having 2 to 6 carbon atoms. The alkynyl group may be substituted or unsubstituted. When substituted, the substituent group(s) is preferably one or more individually selected from halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, thiocarbonyl, xe2x80x94R6NSO2R7, perfluoroalkyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, silyl, ammonium, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycle, heteroaryl and aryl.
A xe2x80x9ccycloalkylxe2x80x9d group refers to an all-carbon monocyclic or fused ring (i.e., rings which share an adjacent pair of carbon atoms) group wherein one of more of the rings does not have a completely conjugated pi-electron system. Examples, without limitation, of cycloalkyl groups are cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane and, cycloheptatriene. A cycloalkyl group may be substituted or unsubstituted. When substituted, the substituent group(s) is preferably one or more individually selected from halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, thiocarbonyl, xe2x80x94R6NSO2R7, perfluoroalkyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, silyl, ammonium, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycle, heteroaryl and aryl.
An xe2x80x9carylxe2x80x9d group refers to an all-carbon monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) groups having a completely conjugated pi-electron system. Examples, without limitation, of aryl groups are phenyl, naphthalenyl and anthracenyl. The aryl group may be substituted or unsubstituted. When substituted, the substituted group(s) is preferably one or more selected halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94RNCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, thiocarbonyl, xe2x80x94R6NSO2R7, perfluoroalkyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, silyl, ammonium, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycle, heteroaryl and aryl.
As used herein, a xe2x80x9cheteroarylxe2x80x9d group refers to a monocyclic or fused ring (i.e., rings which share an adjacent pair of atoms) group having in the ring(s) one or more atoms selected from the group consisting of nitrogen, oxygen and sulfur and, in addition, having a completely conjugated pi-electron system. Examples, without limitation, of heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, purine and carbazole. The heteroaryl group may be substituted or unsubstituted. When substituted, the substituted group(s) is preferably one or more selected from halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CONR6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, thiocarbonyl, xe2x80x94R6NSO2R7, perfluoroalkyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, silyl, ammonium, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycle, heteroaryl and aryl, where X is halogen.
A xe2x80x9cheterocyclic ringxe2x80x9d or xe2x80x9cheterocyclexe2x80x9d group refers to a monocyclic or fused ring group having in the ring(s) one or more atoms selected from the group consisting of nitrogen, oxygen and sulfur. The rings may also have one or more double bonds. However, the rings may or may not have a completely conjugated pi-electron system. The heterocyclic ring may be substituted or unsubstituted. The heterocyci ring may contain one or more oxo groups. When substituted, the substituted group(s) is preferably one or more selected halogen, xe2x80x94OR6, xe2x80x94COR6, xe2x80x94COOR6, OCOR6, xe2x80x94CON6R7, xe2x80x94R6NCOR7, xe2x80x94NR6R7, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CX3, xe2x80x94SR6, xe2x80x94SOR6, xe2x80x94SO2R6, xe2x80x94SO2OR6, xe2x80x94SO2NR6R7, thiocarbonyl, xe2x80x94R6NSO2R7, perfluoroalkyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, silyl, ammonium, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycle, heteroaryl and aryl.
X refers to a halogen group selected from the group consisting of fluorine, chlorine, bromine and iodine.
The definitions of R1-R13 are defined in the present specification.
Compounds that have the same molecular formula but difer in the nature or sequence of bonding of their atoms or arrangements of their atoms in space are termed xe2x80x9cisomers.xe2x80x9d Isomers that differ in the arrangement of their atoms in space are termed xe2x80x9cstereoisomersxe2x80x9d. Stereoisomers that are not mirror images of one another are termed xe2x80x9cdiastereomersxe2x80x9d and those that are non-superimposable mirror images of each other are termed xe2x80x9cenantiomersxe2x80x9d. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or (xe2x88x92)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a xe2x80x9cracemic mixturexe2x80x9d.
The compounds of this invention may possess one or more asymmetric centers; such compounds can therefore be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. For example, if the R6 substituent in a compound of formula (I) is 2-hydroxyethyl, then the carbon to which the hydroxy group is attached is an asymmetric center and therefore the compound of Formula (I) can exist as an (R)- or (S)-stereoisomer. Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art (see discussion in Chapter 4 of xe2x80x9cAdvanced Organic Chemistryxe2x80x9d, 4th edition J. March, John Wiley and Sons, New York, 1992).
The compounds of Formula (I) may exhibit the phenomena of tautomerism and structural isomerism. For example, the compounds described herein may adopt an E or a Z configuration about the double bond connecting the 2-indolinone moiety to the pyrrole moiety or they may be a mixture of E and Z. This invention encompasses any tautomeric or structural isomeric form and mixtures thereof which possess the ability to modulate RTK, CTK and/or STK activity and is not limited to any one tautomeric or structural isomeric form.
It is contemplated that a compound of Formula (I) would be metabolized by enzymes in the body of the organism such as human being to generate a metabolite that can modulate the activity of the protein kinases. Such metabolites are within the scope of the present invention.
As used herein, xe2x80x9cPKxe2x80x9d refers to receptor protein tyrosine kinase (RTKs), non-receptor or xe2x80x9ccellularxe2x80x9d tyrosine kinase (CTKs) and serine-threonine kinases (STKs).
The term xe2x80x9cmethodxe2x80x9d refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by, practitioners of the chemical, pharmaceutical, biological, biochemical and medical arts.
As used herein, the term xe2x80x9cmodulationxe2x80x9d or xe2x80x9cmodulatingxe2x80x9d refers to the alteration of the catalytic activity of RTKs, CTKs and STKs. In particular, modulating refers to the activation of the catalytic activity of RTKs, CTKs and STKs, preferably the activation or inhibition of the catalytic activity of RTKs, CTKs and STKs, depending on the concentration of the compound or salt to which the RTK, CTK or STK is exposed or, more preferably, the inhibition of the catalytic activity of RTKs, CTKs and STKs.
The term xe2x80x9ccatalytic activityxe2x80x9d as used herein refers to the rate of phosphorylation of tyrosine under the influence, direct or indirect, of RTKs and/or CTKs or the phosphorylation of serine and threonine under the influence, direct or indirect, of STKs.
The term xe2x80x9ccontactingxe2x80x9d as used herein refers to bringing a compound of this invention and a target PK together in such a manner that the compound can affect the catalytic activity of the PK, either directly, i.e., by interacting with the kinase itself, or indirectly, i.e., by interacting with another molecule on which the catalytic activity of the kinase is dependent. Such xe2x80x9ccontactingxe2x80x9d can be accomplished in vitro, i.e., in a test tube, a petri dish or the like. In a test tube, contacting may involve only a compound and a PK of interest or it may involve whole cells. Cells may also be maintained or grown in cell culture dishes and contacted with a compound in that environment. In this context, the ability of a particular compound to affect a PK related disorder, i.e., the IC50 of the compound, defined below, can be determined before use of the compounds in vivo with more complex living organisms is attempted. For cells outside the organism, multiple methods exist, and are well-known to those skilled in the art, to get the PKs in contact with the compounds including, but not limited to, direct cell microinjection and numerous transmembrane carrier techniques.
xe2x80x9cIn vitroxe2x80x9d refers to procedures performed in an artificial environment such as, e.g., without limitation, in a test tube or culture medium. The skilled artisan will understand that, for example, an isolated PK may be contacted with a modulator in an in vitro environment. Alternatively, an isolated cell may be contacted with a modulator in an in vitro environment.
As used herein, xe2x80x9cin vivoxe2x80x9d refers to procedures performed within a living organism such as, without limitation, a mouse, rat, rabbit, ungulate, bovine, equine, porcine, canine, feline, primate, or human.
As used herein, xe2x80x9cPK related disorder,xe2x80x9d xe2x80x9cPK driven disorder,xe2x80x9d and xe2x80x9cabnormal PK activityxe2x80x9d all refer to a condition characterized by inappropriate, i.e., under or, more commonly, over, PK catalytic activity, where the particular PK can be an RTK, a CTK or an STK. Inappropriate catalytic activity can arise as the result of either: (1) PK expression in cells which normally do not express PKs, (2) increased PK expression leading to unwanted cell proliferation, differentiation and/or growth, or, (3) decreased PK expression leading to unwanted reductions in cell proliferation, differentiation and/or growth. Over-activity of a PK refers to either amplification of the gene encoding a particular PK or production of a level of PK activity which can correlate with a cell proliferation, differentiation and/or growth disorder (that is, as the level of the PK increases, the severity of one or more of the symptoms of the cellular disorder increases). Under-activity is, of course, the converse, wherein the severity of one or more symptoms of a cellular disorder increase as the level of the PK activity decreases.
As used herein, the terms xe2x80x9cpreventxe2x80x9d, xe2x80x9cpreventingxe2x80x9d and xe2x80x9cpreventionxe2x80x9d refer to a method for barring an organism from acquiring a PK related disorder in the first place.
As used herein, the terms xe2x80x9ctreatxe2x80x9d, xe2x80x9ctreatingxe2x80x9d and xe2x80x9ctreatmentxe2x80x9d refer to a method of alleviating or abrogating a PK mediated cellular disorder and/or its attendant symptoms. With regard particularly to cancer, these terms simply mean that the life expectancy of an individual affected with a cancer will be increased or that one or more of the symptoms of the disease will be reduced.
The term xe2x80x9corganismxe2x80x9d refers to any living entity comprised of at least one cell. A living organism can be as simple as, for example, a single eukaryotic cell or as complex as a mammal. In a preferred aspect, the organism is a mammal. In a particularly preferred aspect, the mammal is a human being.
The term xe2x80x9ctherapeutically effective amountxe2x80x9d as used herein refers to that amount of the compound being administered which will relieve to some extent one or more of the symptoms of the disorder being treated. In reference to the treatment of cancer, a therapeutically effective amount refers to that amount which has the effect of (1) reducing the size of the tumor, (2) inhibiting (that is, slowing to some extent, preferably stopping) tumor metastasis, (3) inhibiting to some extent (that is, slowing to some extent, preferably stopping) tumor growth, and/or, (4) relieving to some extent (or, preferably, eliminating) one or more symptoms associated with the cancer.
By xe2x80x9cmonitoringxe2x80x9d is meant observing or detecting the effect of contacting a compound with a cell expressing a particular PK. The observed or detected effect can be a change in cell phenotype, in the catalytic activity of a PK or a change in the interaction of a PK with a natural binding partner. Techniques for observing or detecting such effects are well-known in the art. For example, the catalytic activity of a PK may be observed by determining the rate or amount of phosphorylation of a target molecule.
xe2x80x9cCell phenotypexe2x80x9d refers to the outward appearance of a cell or tissue or the biological function of the cell or tissue. Examples, without limitation, of a cell phenotype are cell size, cell growth, cell proliferation, cell differentiation, cell survival, apoptosis, and nutrient uptake and use. Such phenotypic characteristics are measurable by techniques well-known in the art.
A xe2x80x9cnatural binding partnerxe2x80x9d refers to a polypeptide that binds to a particular PK in a cell. Natural binding partners can play a role in propagating a signal in a PK-mediated signal transduction process. A change in the interaction of the natural binding partner with the PK can manifest itself as an increased or decreased concentration of the PK/natural binding partner complex and, as a result, in an observable change in the ability of the PK to mediate signal transduction.
As used herein, xe2x80x9cadministerxe2x80x9d or xe2x80x9cadministrationxe2x80x9d refers to the delivery of a compound or salt of the present invention or of a pharmaceutical composition containing a compound or salt of this invention to an organism for the purpose of prevention or treatment of a PK-related disorder.
A xe2x80x9cpharmaceutical compositionxe2x80x9d refers to a mixture of one or more of the compounds described herein, or pharmaceutically acceptable salts or prodrugs thereof, with other chemical components, such as pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.
xe2x80x9cPharmaceutically acceptable excipientxe2x80x9d refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.
xe2x80x9cPharmaceutically acceptable saltxe2x80x9d refers to those salts, which retain the biological effectiveness and properties of the parent compound. Such salts include:
(1) acid addition salt which is obtained by reaction of the free base of the parent compound with inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, and perhcloric acid and the like, or with organic acids such as acetic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, succinic acid or malonic acid and the like, preferably hydrochloric acid or (L)-malic acid; or
(2) salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolanine, tromethamine, N-methylglucamine, and the like.
The compound of Formula (I) may also act as a prodrug. A xe2x80x9cprodrugxe2x80x9d refers to an agent, which is converted into the parent drug in vivo. Prodrugs are often usefull because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent drug is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. An example, without limitation, of a prodrug would be a compound of the present invention, which is, administered as an ester (the xe2x80x9cprodrugxe2x80x9d), carbamate or urea.
The PKs whose catalytic activity is modulated by the compounds of this invention include protein tyrosine kinases of which there are two types, receptor tyrosine kinases (RTKs) and cellular tyrosine kinases (CTKs), and serine-threonine kinases (STKs). RTK mediated signal transduction, is initiated by extracellular interaction with a specific growth factor (ligand), followed by receptor dimerization, transient stimulation of the intrinsic protein tyrosine kinase activity and phosphorylation. Binding sites are thereby created for intracellular signal transduction molecules and lead to the formation of complexes with a spectrum of cytoplasmic signaling molecules that facilitate the appropriate cellular response (e.g., cell division, metabolic effects on the extracellular microenvironment, etc.). See, Schlessinger and Ullrich, Neuron, 9:303-391 (1992).
It has been shown that tyrosine phosphorylation sites on growth factor receptors function as high-affinity binding sites for SH2 (src homology) domains of signaling molecules. Fantl et al., Cell, 69:413-423 (1992), Songyang et al., Mol. Cell. Biol., 14:2777-2785 (1994), Songyang et al., Cell, 72:767-778 (1993), and Koch et al., Science, 252:668-678 (1991). Several intracellular substrate proteins that associate with RTKs have been identified. They may be divided into two principal groups: (1) substrates that have a catalytic domain, and (2) substrates which lack such domain but which serve as adapters and associate with catalytically active molecules. Songyang et al., Cell, 72:767-778 (1993). The specificity of the interactions between receptors and SH2 domains of their substrates is determined by the amino acid residues immediately surrounding the phosphorylated tyrosine residue. Differences in the binding affinities between SH2 domains and the amino acid sequences surrounding the phosphotyrosine residues on particular receptors are consistent with the observed differences in their substrate phosphorylation profiles. Songyang et al., Cell, 72:767-778 (1993). These observations suggest that the function of each RTK is determined not only by its pattern of expression and ligand availability but also by the array of downstream signal transduction pathways that are activated by a particular receptor. Thus, phosphorylation provides an important regulatory step which determines the selectivity of signaling pathways recruited by specific growth factor receptors, as well as differentiation factor receptors.
STKs, being primarily cytosolic, affect the internal biochemistry of the cell, often as a down-line response to a PTK event. STKs have been implicated in the signaling process which initiates DNA synthesis and subsequent mitosis leading to cell proliferation.
Thus, PK signal transduction results in, among other responses, cell proliferation, differentiation, growth and metabolism. Abnormal cell proliferation may result in a wide array of disorders and diseases, including the development of neoplasia such as carcinoma, sarcoma, glioblastoma and hemangioma, disorders such as leukemia, psoriasis, arteriosclerosis, arthritis and diabetic retinopathy and other disorders related to uncontrolled angiogenesis and/or vasculogenesis.
A precise understanding of the mechanism by which the compounds of the invention, in particular, the compounds generated iii vivo from the compounds of the invention, inhibit PKs is not required in order to practice the present invention. However, while not hereby being bound to any particular mechanism or theory, it is believed that the compounds interact with the amino acids in the catalytic region of PKs. PKs typically possess a bi-lobate structure wherein ATP appears to bind in the cleft between the two lobes in a region where the amino acids are conserved among PKs. Inhibitors of PKs are believed to bind by non-covalent interactions such as hydrogen bonding, van der Waals forces and ionic interactions in the same general region where the aforesaid ATP binds to the PKs. More specifically, it is thought that the 2-indolinone component of the compounds of this invention binds in the general space normally occupied by the adenine ring of ATP. Specificity of a particular molecule for a particular PK may then arise as the result of additional interactions between the various substituents on the 2-indolinone core and the amino acid domains specific to particular PKs. Thus, different indolinone substituents may contribute to preferential binding to particular PKs. The ability to select compounds active at different ATP (or other nucleotide) binding sites makes the compounds of this invention useful for targeting any protein with such a site. The compounds disclosed herein may thus have utility as in vitro assays for such proteins as well as exhibiting in vivo therapeutic effects through interaction with such proteins.
In another aspect, the protein kinase, the catalytic activity of which is modulated by contact with a compound of this invention, is a protein tyrosine kinase, more particularly, a receptor protein tyrosine kinase. Among the receptor protein tyrosine kinases whose catalytic activity can be modulated with a compound of this invention, or salt thereof, are, without limitation, selected from the group consisting of Met, Flk, FGFR, PDGFR, HER, IR, IGF, IRR, CSFIR, C-Kit, C-fms, flt. In a preferred aspect, the receptor protein tyrosine kinases whose catalytic activity can be modulated with a compound of this invention, or salt thereof, are, without limitation, Met, Flk-1R, Flk4, KDR/Flk-1, FGFR-1R, FGFR-2R, FGFR-3R, FGFR-4R, PDGFRxcex1, PDGFRxcex2, HER2, HER3, HER4, IRR, CSFIR, C-Kit and C-fms, preferably Met.
The protein tyrosine kinase whose catalytic activity is modulated by contact with a compound of this invention, or a salt thereof, can also be a non-receptor or cellular protein tyrosine kinase (CTK). Thus, the catalytic activity of CTKs such as, without limitation, Src, Frk, Btk, Csk, Abl, ZAP70, Fes, Fps, Fak, Jak, Ack, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr, Aur2 and Yrk may be modulated by contact with a compound or salt of this invention.
Still another group of PKs which may have their catalytic activity modulated by contact with a compound of this invention are the serine-threonine protein kinases such as, without limitation, CDK2, Raf, NEK (including NEK 4a, NEK 4b, NEK 5 and NEK 6) and BUB1.
In another aspect, this invention relates to a method for treating or preventing a PK related disorder by administering a therapeutically effective amount of a compound of this invention, or a salt thereof, to an organism.
It is also an aspect of this invention that a pharmaceutical composition containing a compound of this invention, or a salt thereof, is administered to an organism for the purpose of preventing or treating a PK related disorder.
This invention is therefore directed to compounds that modulate PK signal transduction by affecting the enzymatic activity of RTKs, CTKs and/or STKs, thereby interfering with the signals transduced by such proteins. More particularly, the present invention is directed to compounds which modulate RTK, CTK and/or STK mediated signal transduction pathways as a therapeutic approach to cure many kinds of solid tumors, including but not limited to carcinomas, sarcomas including Kaposi""s sarcoma, erythroblastoma, glioblastoma, meningioma, astrocytoma, melanoma and myoblastoma. Treatment or prevention of non-solid tumor cancers such as leukemia are also contemplated by this invention. Indications may include, but are not limited to brain cancers, bladder cancers, ovarian cancers, gastric cancers, pancreas cancers, colon cancers, blood cancers, lung cancers and bone cancers.
Further examples, without limitation, of the types of disorders related to inappropriate PK activity that the compounds described herein may be useful in preventing, treating and studying, are cell proliferative disorders, fibrotic disorders, metabolic disorders and infectious diseases.
Cell proliferative disorders, which may be prevented, treated or further studied by the present invention include cancer, blood vessel proliferative disorders and mesangial cell proliferative disorders.
Blood vessel proliferative disorders refer to disorders related to abnormal vasculogenesis (blood vessel formation) and angiogenesis (spreading of blood vessels). While vasculogenesis and angiogenesis play important roles in a variety of normal physiological processes such as embryonic development, corpus luteum formation, wound healing and organ regeneration, they also play a pivotal role in cancer development where they result in the formation of new capillaries needed to keep a tumor alive. Other examples of blood vessel proliferation disorders include arthritis, where new capillary blood vessels invade the joint and destroy cartilage, and ocular diseases, like diabetic retinopathy, where new capillaries in the retina invade the vitreous, bleed and cause blindness.
Two structurally related RTKs have been identified to bind VEGF with high affinity: the fms-like tyrosine 1 (fit-1) receptor (Shibuya et al., Oncogene, 5:519-524 (1990); De Vries et al., Science, 255:989-991 (1992) and the KDR/FLK-1 receptor, also known as VEGF-R2. Vascular endothelial growth factor (VEGF) has been reported to be an endothelial cell specific mitogen with in vitro endothelial cell growth promoting activity. Ferrara and Henzel, Biochem. Biophys. Res. Comm., 161:851-858 (1989); Vaisman et al., J. Biol. Chem., 265:19461-19566 (1990). Information set forth in U.S. application Ser. Nos. 08/193,829, 08/038,596 and 07/975,750, strongly suggest that VEGF is not only responsible for endothelial cell proliferation, but also is the prime regulator of normal and pathological angiogenesis. See generally, Klagsbum and Soker, Current Biology, 3(10):699-702 (1993); Houck, et al., J. Biol. Chem., 267:26031-26037 (1992).
Normal vasculogenesis and angiogenesis play important roles in a variety of physiological processes such as embryonic development, wound healing, organ regeneration and female reproductive processes such as follicle development in the corpus luteum during ovulation and placental growth after pregnancy. Folkman and Shing, J. Biological Chem., 267(16):10931-10934 (1992). Uncontrolled vasculogenesis and/or angiogenesis has been associated with diseases such as diabetes as well as with malignant solid tumors that rely on vascularization for growth. Klagsbum and Soker, Current Biology, 3(10):699-702 (1993); Folkham, J. Natl. Cancer Inst., 82:4-6 (1991); Weidner, et al., New Engl. J. Med., 324:1-5 (1991).
As presently understood, the role of VEGF in endothelial cell proliferation and migration during angiogenesis and vasculogenesis indicates an important role for the KDR/FLK-1 receptor in these processes. Diseases such as diabetes mellitus (Folkman, 198, in XIth Congress of Thrombosis and Haemostasis (Verstraeta, et al., eds.), pp. 583-596, Leuven University Press, Leuven) and arthritis, as well as malignant tumor growth may result from uncontrolled angiogenesis. See e.g., Folkman, N. Engl. J. Med., 285:1182-1186 (1971). The receptors to which VEGF specifically binds are an important and powerful therapeutic target for the regulation and modulation of vasculogenesis and/or angiogenesis and a variety of severe diseases which involve abnormal cellular growth caused by such processes. Plowman, et al., DNandP, 7(6):334-339 (1994). More particularly, the KDR/FLK-1 receptor""s highly specific role in neovascularization make it a choice target for therapeutic approaches to the treatment of cancer and other diseases which involve the uncontrolled formation of blood vessels.
Thus, one aspect of the present invention relates to compounds capable of regulating and/or modulating tyrosine kinase signal transduction including KDR/FLK-1 receptor signal transduction in order to inhibit or promote angiogenesis and/or vasculogenesis, that is, compounds that inhibit, prevent, or interfere with the signal transduced by KDR/FLK-1 when activated by ligands such as VEGF. Although it is believed that the compounds of the present invention act on a receptor or other component along the tyrosine kinase signal transduction pathway, they may also act directly on the tumor cells that result from uncontrolled angiogenesis.
Although the nomenclature of the human and murine counterparts of the generic xe2x80x9cflk-Ixe2x80x9d receptor differ, they are, in many respects, interchangeable. The murine receptor, Flk-1, and its human counterpart, KDR, share a sequence homology of 93.4% within the intracellular domain. Likewise, murine FLK-I binds human VEGF with the same affinity as mouse VEGF, and accordingly, is activated by the ligand derived from either species. Millauer et al., Cell, 72:835-846 (1993); Quinn et al., Proc. Natl. Acad. Sci. USA, 90:7533-7537 (1993). FLK-1 also associates with and subsequently tyrosine phosphorylates human RTK substrates (e.g., PLC-xcex3 or p85) when co-expressed in 293 cells (human embryonal kidney fibroblasts).
Models which rely upon the FLK-1 receptor therefore are directly applicable to understanding the KDR receptor. For example, use of the murine FLK-1 receptor in methods which identify compounds that regulate the murine signal transduction pathway are directly applicable to the identification of compounds which may be used to regulate the human signal transduction pathway, that is, which regulate activity related to the KDR receptor. Thus, chemical compounds identified as inhibitors of KDR/FLK-1 in vitro, can be confirmed in suitable in vivo models. Both in vivo mouse and rat animal models have been demonstrated to be of excellent value for the examination of the clinical potential of agents acting on the KDR/FLK-1 induced signal transduction pathway.
Thus, in one aspect, this invention is directed to compounds that regulate, modulate and/or inhibit vasculogenesis and/or angiogenesis by affecting the enzymatic activity of the KDR/FLK-1 receptor and interfering with the signal transduced by KDR/FLK-1. In another aspect, the present invention is directed to compounds which regulate, modulate and/or inhibit the KDR/FLK-1 mediated signal transduction pathway as a therapeutic approach to the treatment of many kinds of solid tumors including, but not limited to, glioblastoma, melanoma and Kaposi""s sarcoma, and ovarian, lung, mammary, prostate, pancreatic, colon and epidermoid carcinoma. In addition, data suggest the administration of compounds which inhibit the KDR/Flk-1 mediated signal transduction pathway may also be used in the treatment of hemangioma, restenois and diabetic retinopathy.
A further aspect of this invention relates to the inhibition of vasculogenesis and angiogenesis by other receptor-mediated pathways, including the pathway comprising the flt-1 receptor.
Receptor tyrosine kinase mediated signal transduction is initiated by extracellular interaction with a specific growth factor (ligand), followed by receptor dimerization, transient stimulation of the intrinsic protein tyrosine kinase activity and autophosphorylation. Binding sites are thereby created for intracellular signal transduction molecules which leads to the formation of complexes with a spectrum of cytoplasmic signaling molecules that facilitate the appropriate cellular response, e.g., cell division and metabolic effects to the extracellular microenvironment. See, Schlessinger and Ullnich, Neuron, 9:1-20 (1992).
The close homology of the intracellular regions of KDR/FLK-1 with that of the PDGF-xcex2 receptor (50.3% homology) and/or the related flt-1 receptor indicates the induction of overlapping signal transduction pathways. For example, for the PDGF-xcex2 receptor, members of the src family (Twamley et al., Proc. Natl. Acad. Sci. USA, 90:7696-7700 (1993)), phosphatidylinositol-3xe2x80x2-kinase (Hu et al., Mol. Cell. Biol., 12:981-990 (1992), phospholipase cxcex3 (Kashishian and Cooper, Mol. Cell. Biol., 4:49-51 (1993)), ras-GTPase-activating protein, (Kashishian et al., EMBO J., 11:1373-1382 (1992), PTP-ID/syp (Kazlauskas et al., Proc. Natl. Acad. Sci. USA, 90:6939-6943 (1993)), Grb2 (Arvidsson et al., Mol. Cell. Biol., 14:6715-6726 (1994)), and the adapter molecules Shc and Nck (Nishimura et al., Mol. Cell. Biol., 13:6889-6896 (1993)), have been shown to bind to regions involving different autophosphorylation sites. See generally, Claesson-Welsh, Prog. Growth Factor Res., 5:37-54 (1994). Thus, it is likely that signal transduction pathways activated by KDR/FLK-1 include the ras pathway (Rozakis et al., Nature, 360:689-692 (1992)), the PI-3xe2x80x2-kinase, the src-mediated and the plcxcex3-mediated pathways. Each of these pathways may play a critical role in the angiogenic and/or vasculogenic effect of KDR/FLK-1 in endothelial cells. Consequently, a still further aspect of this invention relates to the use of the organic compounds described herein to modulate angiogenesis and vasculogenesis as such processes are controlled by these pathways.
Conversely, disorders related to the shrinkage, contraction or closing of blood vessels, such as restenosis, are also implicated and may be treated or prevented by the methods of this invention.
Fibrotic disorders refer to the abnormal formation of extracellular matrices. Examples of fibrotic disorders include hepatic cirrhosis and mesangial cell proliferative disorders. Hepatic cirrhosis is characterized by the increase in extracellular matrix constituents resulting in the formation of a hepatic scar. An increased extracellular matrix resulting in a hepatic scar can also be caused by a viral infection such as hepatitis. Lipocytes appear to play a major role in hepatic cirrhosis. Other fibrotic disorders implicated include atherosclerosis.
Mesangial cell proliferative disorders refer to disorders brought about by abnormal proliferation of mesangial cells. Mesangial proliferative disorders include various human renal diseases such as glomerulonephritis, diabetic nephropathy and malignant nephrosclerosis as well as such disorders as thrombotic microangiopathy syndromes, transplant rejection, and glomerulopathies. The RTK PDGFR has been implicated in the maintenance of mesangial cell proliferation. Floege et al., Kidney International, 43:47S-54S (1993).
Many cancers are cell proliferative disorders and, as noted previously, PKs have been associated with cell proliferative disorders. Thus, it is not surprising that PKs such as, for example, members of the RTK family have been associated with the development of cancer. Some of these receptors, like EGFR (Tuzi et al., Br. J. Cancer, 63:227-233 (1991), Torp et al., APMIS, 100:713-719 (1992)) HER2/neu (Slamon et al., Science, 244:707-712 (1989)) and PDGF-R (Kumabe et al., Oncogene, 7:627-633 (1992)) are over-expressed in many tumors and/or persistently activated by autocrine loops. In fact, in the most common and severe cancers these receptor over-expressions (Akbasak and Suner-Akbasak et al., J. Neurol. Sci., 111:119-133 (1992), Dickson et al., Cancer Treatment Res., 61:249-273 (1992), Korc et al., J. Clin. Invest., 90:1352-1360 (1992)) and autocrine loops (Lee and Donoghue, J. Cell. Biol., 118:1057-1070 (1992), Korc et al., supra, Akbasak and Suner-Akbasak et al., supra) have been demonstrated. For example, EGFR has been associated with squamous cell carcinoma, astrocytoma, glioblastoma, head and neck cancer, lung cancer and bladder cancer. HER2 has been associated with breast, ovarian, gastric, lung, pancreas and bladder cancer. PDGFR has been associated with glioblastoma and melanoma as well as lung, ovarian and prostate cancer. The RTK c-met has also been associated with malignant tumor formation. For example, c-met has been associated with, among other cancers, colorectal, thyroid, pancreatic, gastric and hepatocellular carcinomas and lymphomas. Additionally c-met has been linked to leukemia. Over-expression of the c-met gene has also been detected in patients with Hodgkins disease and Burkitts disease.
IGF-IR, in addition to being implicated in nutritional support and in type-II diabetes, has also been associated with several types of cancers. For example, IGF-I has been implicated as an autocrine growth stimulator for several tumor types, e.g. human breast cancer carcinoma cells (Arteaga et al., J. Clin. Invest., 84:1418-1423 (1989)) and small lung tumor cells (Macauley et al., Cancer Res., 50:2511-2517 (1990)). In addition, IGF-I, while integrally involved in the normal growth and differentiation of the nervous system, also appears to be an autocrine stimulator of human gliomas. Sandberg-Nordqvist et al., Cancer Res., 53:2475-2478 (1993). The importance of IGF-IR and its ligands in cell proliferation is further supported by the fact that many cell types in culture (fibroblasts, epithelial cells, smooth muscle cells, T-lymphocytes, myeloid cells, chondrocytes and osteoblasts (the stem cells of the bone marrow)) are stimulated to grow by IGF-I. Goldring and Goldring, Eukaryotic Gene Expression, 1:301-326 (1991). In a series of recent publications; Baserga suggests that IGF-IR plays a central role in the mechanism of transformation and, as such, could be a preferred target for therapeutic interventions for a broad spectrum of human malignancies. Baserga, Cancer Res., 55:249-252 (1995), Baserga, Cell, 79:927-930 (1994), Coppola et al., Mol. Cell. Biol., 14:4588-4595 (1994).
STKs have been implicated in many types of cancer including, notably, breast cancer (Cance, et al., Int. J. Cancer, 54:571-77 (1993)).
The association between abnormal PK activity and disease is not restricted to cancer. For example, RTKs have been associated with diseases such as psoriasis, diabetes mellitus, endometriosis, angiogenesis, atheromatous plaque development, Alzheimer""s disease, von Hippel-Lindau disease, epidermal hyperproliferation, neurodegenerative diseases, age-related macular degeneration and hemangiomas. For example, EGFR has been indicated in corneal and dermal wound healing. Defects in Insulin-R and IGF-IR are indicated in type-II diabetes mellitus. A more complete correlation between specific RTKs and their therapeutic indications is set forth in Plowman et al., DNandP, 7:334-339 (1994).
As noted previously, not only RTKs but CTKs including, but not limited to, src, abl, fps, yes, fyn, lyn, lck, blk, hck, fgr and yrk (reviewed by Bolen et al., FASEB J., 6:3403-3409 (1992)) are involved in the proliferative and metabolic signal transduction pathway and thus could be expected, and have been shown, to be involved in many PTK-mediated disorders to which the present invention is directed. For example, mutated src (v-src) has been shown to be an oncoprotein (pp60v-src) in chicken. Moreover, its cellular homolog, the proto-oncogene pp60c-src transmits oncogenic signals of many receptors. Over-expression of EGFR or HER2/neu in tumors leads to the constitutive activation of pp60c src, which is characteristic of malignant cells but absent in normal cells. On the other hand, mice deficient in the expression of c-src exhibit an osteopetrotic phenotype, indicating a key participation of c-src in osteoclast function and a possible involvement in related disorders.
Similarly, Zap70 has been implicated in T-cell signaling which may relate to autoimmune disorders.
STKs have been associated with inflammation, autoimmune disease, immunoresponses, and hyperproliferation disorders such as restenosis, fibrosis, psoriasis, osteoarthritis and rheumatoid arthritis.
PKs have also been implicated in embryo implantation. Thus, the compounds of this invention may provide an effective method of preventing such embryo implantation and thereby be useful as birth control agents.
In yet another aspect, the compounds of the instant invention can also be used as anti-infective agents. For example, indolinone compounds are known to exhibit antibacterial and antifungal activities. See, e.g., Singh and Jha, xe2x80x9cIndolinone derivatives as potential antimicrobial agents,xe2x80x9d Zentralbl. Mikrobiol,. 144(2):105-109 (1989). In addition, indolinone compounds have been reported to exhibit significant antiviral activity. See, e.g., Maass et al., xe2x80x9cViral resistance to the thiazolo-iso-indolinones, a new class of nonnucleoside inhibitors of human immunodeficiency virus type 1 reverse transcriptase,xe2x80x9d Antimicrob. Agents Chemother., 37(12):2612-2617 (1993).
Finally, both RTKs and CTKs are currently suspected as being involved in hyperimmune disorders.
A method for identifying a chemical compound that modulates the catalytic activity of one or more of the above discussed protein kinases is another aspect of this invention. The method involves contacting cells expressing the desired protein kinase with a compound of this invention (or its salt) and monitoring the cells for any effect that the compound has on them. The effect may be any observable, either to the naked eye or through the use of instrumentation, change or absence of change in a cell phenotype. The change or absence of change in the cell phenotype monitored may be, for example, without limitation, a change or absence of change in the catalytic activity of the protein kinase in the cells or a change or absence of change in the interaction of the protein kinase with a natural binding partner.
A compound of the present invention or a physiologically acceptable salt thereof, can be administered as such to a human patient or can be administered in pharmaceutical compositions in which the foregoing materials are mixed with suitable carriers or excipient(s). Techniques for formulation and administration of drugs may be found in xe2x80x9cRemington""s Pharmacological Sciences,xe2x80x9d Mack Publishing Co., Easton, Pa., latest edition.
Suitable routes of administration may include, without limitation, oral, intraoral, rectal, transmucosal or intestinal administration or intramuscular, epicutaneous, parenteral, subcutaneous, transdermal, intramedullary, intrathecal, direct intraventricular, intravenous, intravitreal, intraperitoneal, intranasal, intramuscular, intradural, intrarespiratory, nasal inhalation or intraocular injections. The preferred routes of administration are oral and parenteral.
Alternatively, one may administer the compound in a local rather than systemic manner, for example, via injection of the compound directly into a solid tumor, often in a depot or sustained release formulation.
Furthermore, one may administer the drug in a targeted drug delivery system, for example, in a liposome coated with tumor-specific antibody. The liposomes will be targeted to and taken up selectively by the tumor.
Pharmaceutical compositions of the present invention may be manufactured by processes well known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, lyophilizing processes or spray drying.
Pharmaceutical compositions for use in the methods of the present invention may be prepared by any methods of pharmacy, but all methods include the step of bringing in association the active ingredient with the carrier which constitutes one or more necessary ingredients. In particular, pharmaceutical compositions for use in accordance with the present invention may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.
Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, patches, syrups, elixirs, gels, powders, magmas, lozenges, ointments, creams, pastes, plasters, lotions, discs, suppositories, nasal or oral sprays, aerosols and the like.
For injection, the compounds of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such buffers with or without a low concentration of surfactant or cosolvent, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
For oral administration, the compounds can be formulated by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, lozenges, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient. Pharmaceutical preparations for oral use can be made using a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding other suitable auxiliaries if desired, to obtain tablets or dragee cores. Useful excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol, cellulose preparations such as, for example, maize starch, wheat starch, rice starch and potato starch and other materials such as gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinyl-pyrrolidone (PVP). If desired, disintegrating agents may be added, such as cross-linked polyvinyl pyrrolidone, agar, or alginic acid. A salt such as sodium alginate may also be used.
Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopolgel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
Pharmaceutical compositions which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with a filler such as lactose, a binder such as starch, and/or a lubricant such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, liquid polyethylene glycols, cremophor, capmul, medium or long chain mono- di- or triglycerides. Stabilizers may be added in these formulations, also.
For administration by inhalation, the compounds for use according to the present invention are conveniently delivered in the form of an aerosol spray using a pressurized pack or a nebulizer and a suitable propellant, e.g., without limitation, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetra-fluoroethane or carbon dioxide. In the case of a pressurized aerosol, the dosage unit may be controlled by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
The compounds may also be formulated for parenteral administration, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulating materials such as suspending, stabilizing and/or dispersing agents.
Pharmaceutical compositions for parenteral administration include aqueous solutions of a water soluble form, such as, without limitation, a salt, of the active compound. Additionally, suspensions of the active compounds may be prepared in a lipophilic vehicle. Suitable lipophilic vehicles include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate and triglycerides, or materials such as liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers and/or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.
The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, using, e.g., conventional suppository bases such as cocoa butter or other glycerides.
In addition to the formulations described previously, the compounds may also be formulated as depot preparations. Such long acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection. A compound of this invention may be formulated for this route of administration with suitable polymeric or hydrophobic materials (for instance, in an emulsion with a pharmacologically acceptable oil), with ion exchange resins, or as a sparingly soluble derivative such as, without limitation, a sparingly soluble salt.
A non-limiting example of a pharmaceutical carrier for the hydrophobic compounds of the invention is a cosolvent system comprising benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer and an aqueous phase such as the VPD co-solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80(trademark), and 65% w/v polyethylene glycol 300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD:D5W) consists of VPD diluted 1:1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. Naturally, the proportions of such a co-solvent system may be varied considerably without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components may be varied: for example, other low-toxicity nonpolar surfactants may be used instead of Polysorbate 80(trademark), the fraction size of polyethylene glycol may be varied, other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone, and other sugars or polysaccharides may substitute for dextrose.
Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. In addition, certain organic solvents such as dimethylsulfoxide also may be employed, although often at the cost of greater toxicity.
Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.
The pharmaceutical compositions herein also may comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.
Many of the PK modulating compounds of the invention may be provided as physiologically acceptable salts wherein the claimed compound may form the negatively or the positively charged species. Examples of salts in which the compound forms the positively charged moiety include, without limitation, quaternary ammonium (defined elsewhere herein), salts such as the hydrochloride, sulfate, carbonate, lactate, tartrate, maleate, succinate wherein the nitrogen atom of the quaternary ammonium group is a nitrogen of the selected compound of this invention which has reacted with the appropriate acid. Salts in which a compound of this invention forms the negatively charged species include, without limitation, the sodium, potassium, calcium and magnesium salts formed by the reaction of a carboxylic acid group in the compound with an appropriate base (e.g. sodium hydroxide (NaOH), potassium hydroxide (KOH), Calcium hydroxide (Ca(OH)2), etc.).
Dosage
Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an amount sufficient to achieve the intended purpose, i.e., the modulation of PK activity or the treatment or prevention of a PK-related disorder.
More specifically, a therapeutically effective amount means an amount of compound effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated.
Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.
For any compound used in the methods of the invention, the therapeutically effective amount or dose can be estimated initially from cell culture assays. Then, the dosage can be formulated for use in animal models so as to achieve a circulating concentration range that includes the IC50 as determined in cell culture (i.e., the concentration of the test compound which achieves a half-maximal inhibition of the PK activity). Such information can then be used to more accurately determine useful doses in humans.
Toxicity and therapeutic efficacy of the compounds described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the IC50 and the LD50 (both of which are discussed elsewhere herein) for a subject compound. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage may vary depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient""s condition. (See e.g., Fingl, et al., 1975, in xe2x80x9cThe Pharmacological Basis of Therapeuticsxe2x80x9d, Ch. 1 p.1).
Dosage amount and interval may be adjusted individually to provide plasma levels of the active species which are sufficient to maintain the kinase modulating effects. These plasma levels are referred to as minimal effective concentrations (MECs). The MEC will vary for each compound but can be estimated from in vitro data, e.g., the concentration necessary to achieve 50-90% inhibition of a kinase may be ascertained using the assays described herein. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. HPLC assays or bioassays can be used to determine plasma concentrations.
Dosage intervals can also be determined using MEC value. Compounds should be administered using a regimen that maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%. At present, the therapeutically effective amounts of compounds of Formula (I) may range from approximately 25 mg/m2 to 150 mg/m2 perday. Even more preferably 25 mg/m2 to 1000 mg/m2.
In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration and other procedures known in the art may be employed to determine the correct dosage amount and interval.
The amount of a composition administered will, of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.
The compositions may, if desired, be presented in a pack or dispenser device, such as an FDA approved kit, which may contain one or more unit dosage forms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied by a notice associated with the container in a form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the compositions or of human or veterinary administration. Such notice, for example, may be of the labeling approved by the U.S. Food and Drug Administration for prescription drugs or of an approved product insert. Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition. Suitable conditions indicated on the label may include treatment of a tumor, inhibition of angiogenesis, treatment of fibrosis, diabetes, and the like.